Figure 4

Deregulation of the cell cycle by inhibition of Cdk1 and/or Cdk2 boosts expression of NED markers in AR-positive prostate cancer cell lines. A-D, LAPC-4 cells were treated with an inhibitor of Cdk2 activity, CVT-313 (10 μM, 48 h). A, Western blot analysis of Rb protein. B, Analysis of cell cycle distribution. The data represent means ± SD (n=4). C, Western blot analysis of expression of γ-enolase and tubulin β-III. D, qRT-PCR analysis of TUBB3, CHGA, and ENO2 mRNA level. The data represent means ± SD (n=4). “*” denotes statistical significance compared with control. E-G, LAPC-4 cells were transfected with control siRNA or with a combination of Cdk1- and Cdk2 siRNAs for 48 hours. E, Western blot analysis of Cdk1 and Cdk2 48 hours after transfection. F, Analysis of cell cycle distribution in response to Cdk1 and Cdk2 down-regulation. The data represent means ± SD (n=3). G, qRT-PCR analysis of changes in mRNA levels of TUBB3 and CHGA in response to Cdk1 and Cdk2 down-regulation. The data represent means ± SD (n=4). “*”denotes statistical significance compared with control siRNA. H-J, LNCaP and LAPC-4 cells were treated with DMSO or the indicated Cdk1 inhibitors for 48 hours (LNCaP: 2 μM CGP, 7.5 μM RO; LAPC-4: 3.5 μM CGP, 7.5 μM RO). H, Analysis of cell cycle distribution in LNCaP and LAPC-4 cells treated with the indicated Cdk1 inhibitors. The data are presented as means ± SD of one out of two independent experiments performed in duplicate. I, Western blot analysis of γ-enolase and tubulin β-III in response to Cdk1 inhibitors. J, qRT-PCR analysis of TUBB3 and ENO2 in response to Cdk1 inhibitors. The data are presented as means ± SD of one out of two experiments performed in duplicate. CGP, CGP 74514A; RO, RO-3306.